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          胚胎工程大纲
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        <h1 id="胚胎工程"><a href="#胚胎工程" class="headerlink" title="胚胎工程"></a>胚胎工程</h1><h2 id="体内受精和早期胚胎发育"><a href="#体内受精和早期胚胎发育" class="headerlink" title="体内受精和早期胚胎发育"></a>体内受精和早期胚胎发育</h2><h3 id="精子和卵子的发生"><a href="#精子和卵子的发生" class="headerlink" title="精子和卵子的发生"></a>精子和卵子的发生</h3><ul>
<li><p>精子的发生</p>
<ul>
<li>场所：睾丸的曲细精管</li>
<li>时期：初情期到生殖机能衰退</li>
<li>时长：大多家畜两个月左右</li>
<li><p>三个阶段</p>
<ul>
<li>曲细精管壁的精原细胞进行数次有丝分裂，产生大量精原细胞，其中部分精原细胞经过染色体复制和其他物质的合成，进一步形成初级精母细胞。</li>
<li>初级精母细胞连续进行两次分裂（即减数分裂MⅠ和MⅡ）</li>
<li><p>圆形的精子变形（课本P61）</p>
<ul>
<li>细胞核→精子头的主要部分</li>
<li>高尔基体→头部的顶体</li>
<li>中心体→精子的尾</li>
<li>线粒体→线粒体鞘</li>
<li>其他物质浓缩为球状，叫做原生质滴，直到最后脱落</li>
</ul>
</li>
</ul>
</li>
<li><p>PS：不同动物的精子形态相似，大小略有不同，但与动物体型大小无关。</p>
</li>
</ul>
</li>
<li><p>卵子的发生</p>
<ul>
<li>场所：卵巢</li>
<li>时期：胚胎性别分化以后</li>
<li><p>过程（书P62-P63）</p>
<ul>
<li>性别分化以后，雌性胎儿卵巢内的卵原细胞有丝分裂增加数量，演变为初级卵母细胞，这时，它被卵泡细胞包围，形成卵泡。</li>
<li><p>排卵前后完成减数第一次分裂，产生一个次级卵母细胞和第一极体，进入输卵管，准备与精子受精。</p>
<ul>
<li>PS：透明带在初情期刚开始形成</li>
</ul>
</li>
<li><p>精子与卵子结合的过程中进行减数第二次分裂，次级卵母细胞经分裂产生一个成熟的卵子和第二极体</p>
<ul>
<li>当在卵细胞膜和透明带的间隙可以观察到两个极体时，说明卵子已经完成了受精，这是判断卵子是否受精的标志。</li>
</ul>
</li>
</ul>
</li>
<li><p>PS：哺乳动物卵泡的形成和在卵巢内的储备，是在出生前（胎儿时期）完成的</p>
</li>
<li>PS：多数哺乳动物减数第一次分裂产生的第一极体不再分裂</li>
</ul>
</li>
</ul>
<h3 id="受精"><a href="#受精" class="headerlink" title="受精"></a>受精</h3><ul>
<li>概念：精子与卵子结合形成合子的过程</li>
<li>场所：雌性动物输卵管</li>
<li><p>阶段</p>
<ul>
<li><p>准备阶段</p>
<ul>
<li><p>精子获能</p>
<ul>
<li>精子必须在雌性动物生殖道发生相应生理变化后才能获得受精能力</li>
</ul>
</li>
<li><p>卵子的准备</p>
<ul>
<li>一般排出后 2~3 h才能被精子穿入</li>
<li>要经历类似于精子获能的过程</li>
<li>在输卵管内进一步成熟，达到减数第二次分裂的中期时，才具备与精子受精的能力</li>
</ul>
</li>
</ul>
</li>
<li><p>受精阶段（书P65）</p>
<ul>
<li><p>精子穿越放射冠</p>
<ul>
<li><p>顶体反应</p>
<ul>
<li>获能后的精子释放顶体酶，溶解卵丘细胞之间的物质，形成精子穿越放射冠的通路</li>
</ul>
</li>
</ul>
</li>
<li><p>精子穿越透明带</p>
<ul>
<li>顶体酶将透明带溶出一条孔道</li>
<li><p>透明带反应</p>
<ul>
<li>是防止多个精子进入透明带，引起多精入卵受精的第一道屏障</li>
</ul>
</li>
</ul>
</li>
<li><p>精子进入卵细胞膜</p>
<ul>
<li>卵细胞膜表面有大量微绒毛，精子与卵细胞膜接触时，立即被微绒毛抱合，随后，精子外膜与卵细胞膜相互融合，精子入卵</li>
<li><p>卵细胞膜反应</p>
<ul>
<li>第二道屏障</li>
</ul>
</li>
</ul>
</li>
<li><p>原核形成和融合</p>
<ul>
<li>精子入卵后，尾部脱离，原有核膜破裂，形成一个新的核膜，最后形成一个比原来精子核还大的核，叫做雄原核</li>
<li>被激活的卵子完成减数第二次分裂，排出第二极体后，形成雌原核</li>
<li>PS：雌原核一般略小于雄原核</li>
<li>雌雄原核充分发育后，相向移动，彼此接触，二者体积缩小、合并，形成合子</li>
</ul>
</li>
</ul>
</li>
</ul>
</li>
</ul>
<h3 id="胚胎发育"><a href="#胚胎发育" class="headerlink" title="胚胎发育"></a>胚胎发育</h3><ul>
<li>合子形成后即在输卵管中进行有丝分裂，开始发育</li>
<li><p>阶段（书P66）</p>
<ul>
<li><p>卵裂期</p>
<ul>
<li>在透明带内进行，总体积不增加或略有缩小</li>
</ul>
</li>
<li><p>桑椹胚（实际属于卵裂期）</p>
<ul>
<li>胚胎细胞32个左右</li>
<li>每一个细胞都是全能细胞</li>
</ul>
</li>
<li><p>囊胚</p>
<ul>
<li>开始分化</li>
<li><p>结构</p>
<ul>
<li><p>内细胞团</p>
<ul>
<li>发育成胎儿的各种组织</li>
</ul>
</li>
<li><p>滋养层</p>
<ul>
<li>发育成胎膜和胎盘</li>
</ul>
</li>
<li><p>囊胚腔</p>
</li>
</ul>
</li>
<li><p>孵化</p>
<ul>
<li>囊胚扩大，导致透明带破裂，胚胎从其中伸展出来</li>
</ul>
</li>
</ul>
</li>
<li><p>原肠胚</p>
<ul>
<li><p>结构</p>
<ul>
<li>滋养层</li>
<li><p>外胚层</p>
<ul>
<li>内细胞团表层细胞</li>
</ul>
</li>
<li><p>内胚层</p>
<ul>
<li>内细胞团下方细胞</li>
</ul>
</li>
<li><p>原肠腔</p>
</li>
</ul>
</li>
</ul>
</li>
</ul>
</li>
</ul>
<h2 id="体外受精和早期胚胎培养"><a href="#体外受精和早期胚胎培养" class="headerlink" title="体外受精和早期胚胎培养"></a>体外受精和早期胚胎培养</h2><h3 id="体外受精"><a href="#体外受精" class="headerlink" title="体外受精"></a>体外受精</h3><ul>
<li><p>卵母细胞的采集和培养</p>
<ul>
<li><p>对实验动物（小鼠、兔），以及家畜（猪、羊）</p>
<ul>
<li>促性腺激素超数排卵处理</li>
<li>从输卵管中冲取卵子</li>
<li>直接与获能的精子受精</li>
</ul>
</li>
<li><p>对大家畜或大型动物（牛）</p>
<ul>
<li><p>主要方法</p>
<ul>
<li>屠宰场卵巢</li>
<li>活体采卵</li>
</ul>
</li>
<li><p>采集的卵母细胞都要经人工体外培养成熟</p>
</li>
</ul>
</li>
</ul>
</li>
<li><p>精子的采集和获能</p>
<ul>
<li><p>采集</p>
<ul>
<li>假阴道法</li>
<li><p>手握法</p>
<ul>
<li>只适用于小型家畜</li>
</ul>
</li>
<li><p>电刺激法</p>
<ul>
<li>适用于野生或经济动物</li>
<li>刺激腰荐部射精中枢神经</li>
</ul>
</li>
</ul>
</li>
<li><p>获能处理</p>
<ul>
<li><p>对啮齿动物、家兔、猪</p>
<ul>
<li>培养法：将取自附睾的精子放入人工配制的获能液中</li>
</ul>
</li>
<li><p>对牛、羊等家畜</p>
<ul>
<li><p>化学诱导法</p>
<ul>
<li>肝素溶液</li>
<li>钙离子载体A23187溶液</li>
</ul>
</li>
</ul>
</li>
</ul>
</li>
</ul>
</li>
<li><p>受精</p>
<ul>
<li>场所：获能溶液或专用的受精溶液</li>
<li>一般要放在培养液小滴中共同培养一段时间才能完成受精</li>
<li>精子获能前离心处理——精清中有物质会抑制</li>
</ul>
</li>
</ul>
<h3 id="胚胎的早期培养"><a href="#胚胎的早期培养" class="headerlink" title="胚胎的早期培养"></a>胚胎的早期培养</h3><ul>
<li>体外受精后将受精卵移入发育培养液</li>
<li><p>培养液成分</p>
<ul>
<li>无机盐、有机盐</li>
<li>维生素、激素</li>
<li>氨基酸、核苷酸</li>
<li>血清</li>
</ul>
</li>
<li><p>发育到适宜阶段时可移植可冷冻保存</p>
</li>
<li><p>PS：不同动物移植时间不同</p>
<ul>
<li>牛、羊——桑椹胚、囊胚</li>
<li>小鼠家兔较上者更早</li>
<li>人—— 8~16 个细胞阶段</li>
</ul>
</li>
</ul>
<h2 id="胚胎工程的应用及前景"><a href="#胚胎工程的应用及前景" class="headerlink" title="胚胎工程的应用及前景"></a>胚胎工程的应用及前景</h2><h3 id="胚胎移植"><a href="#胚胎移植" class="headerlink" title="胚胎移植"></a>胚胎移植</h3><ul>
<li>概念</li>
<li>现状和意义</li>
<li><p>生理学基础</p>
<ul>
<li>哺乳动物发情排卵后，不管是否妊娠，在一段时间内，同种动物的供、受体生殖器官的生理变化是相同的</li>
<li><p>哺乳动物的早期胚胎形成后，在一定时间内不会与母体子宫建立组织上的联系，而是处于游离状态</p>
<ul>
<li>为胚胎收集提供可能</li>
</ul>
</li>
<li><p>受体对移入子宫的外来胚胎不产生免疫排斥反应</p>
</li>
<li>供体胚胎可与受体子宫建立正常的生理和组织联系，但遗传特性不受任何影响</li>
</ul>
</li>
<li><p>基本程序</p>
<ul>
<li>①用激素对供、受体同期发情</li>
<li>②供体超数排卵</li>
<li>③人工受精</li>
<li><p>④‘胚胎收集</p>
<ul>
<li>在配种或输精后第 7 天</li>
<li>用特殊冲卵装置冲卵</li>
</ul>
</li>
<li><p>⑤胚胎质量检查</p>
<ul>
<li>应为桑椹胚或囊胚</li>
</ul>
</li>
<li><p>⑥胚胎移植或 -196℃ 液氮保存</p>
</li>
<li><p>⑦胚胎的移植</p>
<ul>
<li><p>手术法</p>
<ul>
<li>引出受体子宫和卵巢，将胚胎注入子宫角</li>
</ul>
</li>
<li><p>非手术法</p>
<ul>
<li>移植管送入子宫相应部位</li>
</ul>
</li>
</ul>
</li>
<li><p>⑧进行是否妊娠的检查</p>
</li>
</ul>
</li>
</ul>
<h3 id="胚胎分割"><a href="#胚胎分割" class="headerlink" title="胚胎分割"></a>胚胎分割</h3><ul>
<li>概念</li>
<li><p>主要仪器设备</p>
<ul>
<li>实体显微镜</li>
<li>显微操作仪</li>
</ul>
</li>
<li><p>选择桑椹胚或囊胚，用分割针或分割刀进行分割</p>
</li>
<li><p>具体操作</p>
<ul>
<li>用分割针或分割刀将胚胎切开</li>
<li>吸出半个胚胎注入预先准备的空透明带，或直接将裸半胚移入受体</li>
<li>PS：囊胚的话，内细胞团要均等分割</li>
<li>PPS：取样滋养层可以做DNA分析性别鉴定</li>
</ul>
</li>
</ul>
<h3 id="胚胎干细胞"><a href="#胚胎干细胞" class="headerlink" title="胚胎干细胞"></a>胚胎干细胞</h3><ul>
<li>概念：又称 ES 或 EK细胞，是由早期胚胎或原始性腺中分离出来的一类细胞</li>
<li><p>特点</p>
<ul>
<li><p>具有胚胎细胞的特性</p>
<ul>
<li><p>形态上</p>
<ul>
<li>体积小</li>
<li>细胞核大</li>
<li>核仁明显</li>
</ul>
</li>
<li><p>功能上</p>
<ul>
<li>具有发育的全能性</li>
</ul>
</li>
</ul>
</li>
<li><p>在体外培养条件下可以增殖而不分化</p>
</li>
<li>可以冷冻保存，也可以遗传改造</li>
</ul>
</li>
<li><p>应用</p>
<ul>
<li>治疗某些顽症</li>
</ul>
</li>
<li><p>是研究体外细胞分化的理想材料</p>
<ul>
<li>在饲养层细胞（一般为输卵管上皮细胞）上，或添加了抑制因子的培养液中，能够维持不分化的状态</li>
<li>加入分化诱导因子（如：牛磺酸、丁酰环腺苷酸），可以诱导 ES 细胞向不同组织细胞分化</li>
</ul>
</li>
</ul>

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          <div class="post-toc motion-element"><ol class="nav"><li class="nav-item nav-level-1"><a class="nav-link" href="#胚胎工程"><span class="nav-number">1.</span> <span class="nav-text">胚胎工程</span></a><ol class="nav-child"><li class="nav-item nav-level-2"><a class="nav-link" href="#体内受精和早期胚胎发育"><span class="nav-number">1.1.</span> <span class="nav-text">体内受精和早期胚胎发育</span></a><ol class="nav-child"><li class="nav-item nav-level-3"><a class="nav-link" href="#精子和卵子的发生"><span class="nav-number">1.1.1.</span> <span class="nav-text">精子和卵子的发生</span></a></li><li class="nav-item nav-level-3"><a class="nav-link" href="#受精"><span class="nav-number">1.1.2.</span> <span class="nav-text">受精</span></a></li><li class="nav-item nav-level-3"><a class="nav-link" href="#胚胎发育"><span class="nav-number">1.1.3.</span> <span class="nav-text">胚胎发育</span></a></li></ol></li><li class="nav-item nav-level-2"><a class="nav-link" href="#体外受精和早期胚胎培养"><span class="nav-number">1.2.</span> <span class="nav-text">体外受精和早期胚胎培养</span></a><ol class="nav-child"><li class="nav-item nav-level-3"><a class="nav-link" href="#体外受精"><span class="nav-number">1.2.1.</span> <span class="nav-text">体外受精</span></a></li><li class="nav-item nav-level-3"><a class="nav-link" href="#胚胎的早期培养"><span class="nav-number">1.2.2.</span> <span class="nav-text">胚胎的早期培养</span></a></li></ol></li><li class="nav-item nav-level-2"><a class="nav-link" href="#胚胎工程的应用及前景"><span class="nav-number">1.3.</span> <span class="nav-text">胚胎工程的应用及前景</span></a><ol class="nav-child"><li class="nav-item nav-level-3"><a class="nav-link" href="#胚胎移植"><span class="nav-number">1.3.1.</span> <span class="nav-text">胚胎移植</span></a></li><li class="nav-item nav-level-3"><a class="nav-link" href="#胚胎分割"><span class="nav-number">1.3.2.</span> <span class="nav-text">胚胎分割</span></a></li><li class="nav-item nav-level-3"><a class="nav-link" href="#胚胎干细胞"><span class="nav-number">1.3.3.</span> <span class="nav-text">胚胎干细胞</span></a></li></ol></li></ol></li></ol></div>
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